The CBLL1 Knockout SK-HEP-1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout population of the human SK-HEP-1 hepatocellular carcinoma cell line, enabling loss-of-function analysis of the CBLL1 (HAKAI) gene. This polyclonal pool, generated via CRISPR/Cas9-mediated gene disruption, offers a heterogeneous knockout model suitable for studying gene function without clonal selection bias.
SK-HEP-1 is an ascites-derived liver adenocarcinoma cell line that retains epithelial characteristics and serves as a well-established model for hepatocellular carcinoma (HCC) research. The cells exhibit active EMT, Wnt, and TGF-beta signaling pathways, which are central to hepatic carcinogenesis and metastasis.
CBLL1 encodes an E3 ubiquitin ligase that ubiquitinates E-cadherin, marking it for endocytic degradation and thus disrupting adherens junctions. This process is triggered by TGF-beta, Wnt ligands, and Src kinase, leading to the release of beta-catenin from the membrane. Stabilized beta-catenin translocates to the nucleus, where it partners with TCF/LEF transcription factors to drive expression of Wnt target genes. CBLL1 also interacts with c-Met and Grb2, integrating signals from growth factor receptors. Consequently, CBLL1 promotes epithelial?Cmesenchymal transition (EMT), migration, and invasion.
In the SK-HEP-1 context, CBLL1 knockout is particularly relevant for dissecting the molecular switches governing HCC malignancy. Loss of CBLL1 function in these cells can dampen E-cadherin degradation, potentially restoring cell adhesion and attenuating Wnt/beta-catenin signaling. The polyclonal nature of the knockout population reflects heterogeneous responses akin to those in tumor tissue, providing a robust system to evaluate CBLL1’s impact on cancer cell behavior.
Typical applications include western blotting to assess E-cadherin and beta-catenin protein levels, Boyden chamber or scratch assays to measure migration and invasion, and TCF/LEF luciferase reporters to gauge Wnt pathway activity. Co-immunoprecipitation experiments can verify disrupted CBLL1?CE-cadherin complexes, while ubiquitination assays directly monitor E-cadherin modification status. Immunofluorescence can reveal rescued E-cadherin localization at cell?Ccell contacts, and RT-qPCR can profile changes in EMT markers. These polyclonal CBLL1 knockout SK-HEP-1 cells offer a versatile platform for investigating CBLL1 function in hepatocellular carcinoma and EMT. For additional information, please contact Ascent Research.
